| 摘要: |
| 为了增强单个蛋白的抗原性,利用(Gly4Ser)3柔性连接肽将牛结核杆菌MPB70和ESAT-6融合。采用重叠延伸PCR技术将牛结核杆菌mpb70与esat-6基因连接,获得了融合基因mpb70-esat-6,连接至T-Vector pMD19中,获得了克隆质粒pMD-70-esat-6。经BamH I、EcoR I酶切、纯化,并与 pET28a(+)载体连接,构建了pET-70-esat-6重组表达质粒。SDS-PAGE发现,在27.2ku处表达了融合蛋白MPB70-ESAT-6,Western blotting证实,MPB70-ESAT-6与牛结核阳性血清反应性良好。为研究MPB70-ESAT-6融合蛋白作为牛结核病的诊断抗原及疫苗奠定了基础。 |
| 关键词: 牛结核杆菌 mpb70基因 esat-6基因 融合蛋白 原核表达 |
| DOI： |
| 投稿时间：2019-01-11修订日期：2019-03-07 |
| 基金项目:国家自然科学(No.31572555)；吉林省科技发展计划项目(No.20170101025JC)；吉林省教育厅科学技术研究项目(No.201441) |
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| Prokaryotic Expression of MPB70-ESAT-6 Fusion Protein of Mycobacterium bovis and its Antigen reactivity Analysis |
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| (Jilin Agricultural University) |
| Abstract: |
| To enhance the antigenicity of individual protein, Mycobacterium bovis(M. bovis) strain MPB70 was fused with ESAT-6 via a flexible linker, (Gly4Ser)3. The fusion gene mpb70-esat-6 was obtained using overlap extension PCR and linked to pMD19 T-Vector. After BamH I and EcoR I restriction enzyme digestion, the cloned plasmid pMD-70-esat-6 was connected with pET28a(+) vector to construct a recombinat plasmid pET-70-esat-6. SDS-PAGE showed that fusion protein mpb70-esat-6 was expressed at 27.2ku. MPB70-ESAT-6 exhibited good reactivity with M. bovis positive serum confirmed by Western |
| Key words: Mycobacterium bovis mpb70 gene esat-6 gene fusion protein Prokaryotic expression |
